- Volume 49, Issue 2, 1999

Phenotypic and phylogenetic studies were performed on a hitherto undescribed micro-organism isolated from a minke whale (Balaenoptera acutorostrata). Comparative 16S rRNA gene sequencing studies demonstrated that the unknown strain constituted a new subline close to, but distinct from, Abiotrophia adiacens and Abiotrophia elegans. The unknown bacterium was readily distinguished from these two Abiotrophia species by biochemical tests and electrophoretic analysis of whole-cell proteins. On the basis of phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Abiotrophia balaenopterae sp. nov., the type strain of which is M1975/96/1T (= CCUG 37380T).

An almost complete sequence of the 16S rDNA of the type strain of Amycolatopsis rugosa was determined following direct sequencing of the amplified gene. The sequence was aligned with those of representatives of the family Pseudonocardiaceae and related actinomycetes and phylogenetic trees were inferred by using three tree-making algorithms. The organism formed a distinct clade within the evolutionary radiation occupied by the family Pseudonocardiaceae. It was also readily distinguished from all of the validly described genera classified in this taxon by using a combination of chemical and morphological markers. On the basis of these genotypic and phenotypic differences, the name Prauserella gen. nov. is proposed for a new genus containing the previously misclassified Amycolatopsis rugosa. The type strain of Prauserella rugosa is DSM 43194T (= ATCC 43014T = NCIMB 8926T).

A group of 91 moderately halophilic, Gram-positive, rod-shaped strains were isolated from enrichments prepared from Dead Sea water samples collected 57 years ago. These strains were examined for 117 morphological, physiological, biochemical, nutritional and antibiotic susceptibility characteristics. All strains formed endospores and were motile, strictly aerobic and positive for catalase and oxidase. They grew in media containing 5–25% (w/v) total salts, showing optimal growth at 10% (w/v). Eighteen strains were chosen as representative isolates and were studied in more detail. All these strains had meso-diaminopimelic acid in the cell wall and a DNA G+C content of 39·0–42·8 mol%; they constitute a group with levels of DNA-DNA similarity of 70-100%. The sequences of the 16S rRNA genes of three representative strains (strains 123T, 557 and 832) were almost identical (99·9%), and placed the strains in the low G+C content Gram-positive bacteria. On the basis of their features, these isolates should be regarded as members of a new species of the genus Bacillus, for which the name Bacillus marismortui sp. nov. is proposed. The type strain is strain 123T (= DSM 12325T = ATCC 700626T = CIP 105609T = CECT 5066T).

Almost complete 16S rRNA gene sequences were generated for the type strains of the obligate insect pathogens Bacillus lentimorbus and Bacillus popilliae and a second strain of Bacillus popilliae (NRRL B-4081) received as ‘Bacillus popilliae var. melolonthae’. A phylogenetic tree was constructed which grouped these strains into a well defined subcluster within the genus Paenibacillus. Bacillus popilliae NRRL B-4081 occupied an intermediate position between the type strains of Bacillus lentimorbus and Bacillus popilliae but with a marked clustering to the latter. The phylogenetic assignment of these strains to Paenibacillus is in contrast to earlier studies which placed these bacteria in the genus Bacillus, close to Bacillus subtilis. Indeed, the rRNA sequences generated in this study share less than 88% similarity to the deposited sequences for Bacillus popilliae ATCC 14706T and Bacillus lentimorbus ATCC 14707T. The results obtained by using different tree algorithms, bootstrap analysis, branch lengths and verification by signature nucleotide analysis supported the reclassif ication of these species in the genus Paenibacillus as Paenibacillus lentimorbus comb. nov. and Paenibacillus popilliae comb. nov.

Three strains of Eubacterium aerofaciens, JCM 10188T, JCM 7790 and JCM 7791, and 178 freshly isolated strains of the Eubacterium aerofaciens group from human faeces were characterized by biochemical tests, cell wall peptidoglycan type and 16S rRNA analysis. The Eubacterium aerofaciens group was divided into four groups by fermentation patterns of sucrose and cellobiose, and were further divided into 16 sub-groups by fermentation patterns of aesculin, salicin and amygdalin. All of the strains of the Eubacterium aerofaciens group were shown to be phylogenetically distantly related to Eubacterium limosum, which is the type species of genus Eubacterium. Eubacterium aerofaciens was shown to have a specific phylogenetic association with Coriobacterium glomerans. All the strains belonging to Eubacterium aerofaciens resembled Coriobacterium glomerans in possessing a high G+C content (60 mol%). Cell wall analysis, however, revealed the presence of different A4β (l-Ala)-d-Glu-l-Orn-l-Asp peptidoglycan types. Based on a 16S rRNA sequence divergence of greater than 9% with Coriobacterium glomerans and the presence of a unique peptidoglycan type, a new genus, Collinsella, is proposed for Eubacterium aerofaciens, with one species, Collinsella aerofaciens. The type strain of Collinsella aerofaciens is JCM 10188T.

16S rRNA gene sequences were determined for Eubacterium exiguum and Peptostreptococcus heliotrinreducens. These species were found to be closely related and, together with Eubacterium lentum, to constitute a branch of the Coriobacteriaceae. Two new genera are proposed on the basis of phenotypic characteristics and 16S rRNA gene sequence comparisons: Slackia to include the bile-sensitive species Eubacterium exiguum and P. heliotrinreducens, and Eggerthella to include the bile-resistant Eubacterium lentum. It is proposed that Eubacterium exiguum and Peptostreptococcus heliotrinreducens are transferred to the genus Slackia gen. nov. as Slackia exigua gen. nov., comb. nov. (type strain ATCC 700122T) and Slackia heliotrinireducens gen. nov., comb. nov. (type strain NTCC 11029T). respectively, and Eubacterium lentum is transferred to the genus Eggerthella gen. nov. as Eggerthella lenta gen. nov., comb. nov. with Eggerthella lenta as the type species.

A novel extremely thermophilic bacterium was isolated from the world's deepest sea-floor, the Mariana Trench Challenger Deep at a depth of 10897 m. Cells were Gram-reaction variable, non-spore-forming and non-motile rods without flagella. Growth was observed between 50 and 80 °C (optimum: 74–76 C; 90 min doubling time), pH 5·4 and 9·5 (optimum: pH 7·0–7·5) and 0·5 and 5% sea salts (optimum: 2 % sea salts). The isolate was a strictly aerobic heterotroph capable of utilizing as sole energy and carbon source: yeast extract, peptone, cellulose, starch, chitin, casein, Casamino acids, a variety of sugars, carboxylic acids and amino acids. The G+C content of the genomic DNA was 72·5 mol%. Phylogenetic analysis based on 16S rRNA sequences placed this aerobic, high-G+C-content bacterium among the members of the Gram-positive, low-G+C-content anaerobic thermophilic bacteria within the Bacillus-Clostridium subphylum. On the basis of the physiological and molecular properties of the new isolate, the name Thermaerobacter marianensis gen. nov., sp. nov. (type strain 7p75aT = JCM 10246T) is proposed.

Two strains of a Gram-positive catalase-negative, facultatively anaerobic coccus originating from human sources were characterized by phenotypic and molecular taxonomic methods. The strains were found to be identical to each other based on 16S rRNA gene sequencing and constitute a new subline within the genus Facklamia. The unknown bacterium was readily distinguished from Facklamis hominis and Facklamia ignava by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence it is proposed that the unknown bacterium be classified as Facklamia sourekii sp. nov., the type strain of which is CCUG 28783AT.

Saccharomonospora species were subjected to DNA-DNA relatedness determination to investigate their genetic relationships based on genomic DNA. The levels of DNA-DNA relatedness among the type strains of the four validly described Saccharomonospora species ranged from 18 to 46%. Saccharomonospora viridis KCTC 9156. which was described as ′Saccharomonospora internatus′ but later reclassified as a member of S. viridis, showed a similarity level of 90% to S. viridis KCTC 9115T. Saccharomonospora sp. strain K180 exhibited levels of DNA-DNA relatedness that are indicative of a new species. Saccharomonospora azurea exhibited levels of DNA-DNA homology of 84–95% to ′Saccharomonospora caesia′ strains including the type strain. Therefore, on the basis of DNA-DNA relatedness data and taxonomic data described previously, it is proposed that ′S. caesia′ should be considered as a synonym of S. azurea.

A p-nitrophenol-degrading bacterial strain was isolated from industrial wastewater. This strain (NSP41T) was identified as a member of the genus Nocardioides from chemotaxonomic characterizations and phylogenetic inference based on 16S rDNA sequence analysis. The G+C content is 71·4 mol%. The diamino acid in the cell-wall peptidoglycan is ll-diaminopimelic acid. The predominant menaquinone is MK-8(H4). The cellular fatty acid profile is similar to those of Nocardioides species. 16S rDNA sequence analysis show that strain NSP41T is the most related to Nocardioides simplex strains with a level of nucleotide similarity of 98·6%. The levels of 16S rDNA similarity between strain NSP41T and other Nocardioides species ranged from 93·8 to 95·1 %. This organism is distinguishable from some other Nocardioides species as well as N. simplex strains by DNA–DNA relatedness data. This organism is different from N. simplex strains in some phenotypic characteristics. Therefore, on the basis of the data presented, a new species of the genus Nocardioides, Nocardioides nitrophenolicus, is proposed. The type strain of the new species is strain NSP41T ( = KCTC 0457BPT).

The taxonomic status of an actinomycete (MA140/96T) isolated from indoor building materials of a children’s day care centre was studied using the polyphasic approach. The cell morphology was atypical for an actinomycete, electron microscopy revealed a hairy surface, highly unusual for Gram-positive bacteria. The organisms grew at 10–37 °C, no growth was visible at 5 °C and 45 °C in 5 d. The cell wall contained the diamino acid meso-diaminopimelic acid and the sugars arabinose, galactose, mannose and ribose. The phospholipids phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol were detected. The only menaquinone found was MK-9(H2). The fatty acid pattern was composed of palmitic acid (23·6%) palmitoleic acid (16·5%) and another hexadecenoic acid 16:1cis11 (1·4%), oleic acid (29·9%), stearic acid (2·9%) and the 10-methyl-branched tuberculostearic acid (23·2%). A gas-chromatographic analysis of the mycolic acid revealed a carbon-chain length of C50–C56. The G+C was 64·8 mol%. The results of 16S rDNA sequence comparisons revealed that strain MA140/96T represents a new lineage in the suborder Corynebacterineae of the order Actinomycetales. Therefore, it was concluded that strain MA140/96T should be assigned to a new genus and species, for which the name Williamsia muralis gen. nov., sp. nov. is proposed. The type strain of the species is MA140/96T (= DSM 44343T).

The one-dimensional whole-cell protein patterns and a variety of biochemical characteristics of the type and reference strains of Streptococcus intestinalis and Streptococcus alactolyticus were studied. All the data revealed that strain of both species were indistinguishable. Reported differences in haemolytic activity and presence of Lancefield antigens were not reproduced. All strains were α-haemolytic. The S. intestinalis type strain possessed the group G antigen, but a second S. intestinalis reference strain and all of the S. alactolyticus strains possessed the group D antigen. All strains produced urease activity (albeit some after prolonged incubation), a characteristic considered important for S. intestinalis. Given the congruence between wholecell protein electrophoresis and percentage of DNA-DNA hybridization, these data suggest that Streptococcus intestinalis Robinson et al. 1988 is a junior synonym of Streptococcus alactolyticus Farrow et al. 1984.

By 16S rDNA sequence analysis, two strains identified previously as members of the genus Nocardioides, namely ‘Nocardioides fulvus’ IFO 14399 and Nocardioides sp. ATCC 39419, were found to form lines of descent distinct from the genus Nocardioides. Therefore, the two strains were studied taxonomically in detail by phenotypic, particularly chemotaxonomic, characterization, and by phylogenetic analysis including other related taxa. The genomic DNA G+C contents of ‘N. fulvus’ IFO 14399 and Nocardioides sp. ATCC 39419 are 70 and 68 mol%, respectively. The two strains contain ll-diaminopimelic acid as the diamino acid in their peptidoglycan, together with l-alanine at position 1 of the peptide subunit. The predominant menaquinone found in the two strains is MK-9(H4). The major cellular fatty acid found in the two strains is anteiso-C15:0, which is a minor component of members of the genus Nocardioides. The diagnostic polar lipid of the two strains is phosphatidylcholine. A phylogenetic tree was constructed which showed that the two strains form evolutionary lineages distinct from other ll-diaminopimelic-acid-containing taxa such as, for example, the genus Nocardioides. On the basis of our data, a new genus, Kribbella gen. nov., and two new species, Kribbella flavida gen. nov., sp. nov. and Kribbellla sandramycini gen. nov., sp. nov., are proposed. It is also proposed that the new genus be included in the family Nocardioidaceae.

An actinomycete, strain YC75T, which produced bafilomycin-like antifungal compounds, was identified as a member of the genus Kitasatospora on the basis of morphological and chemotaxonomic characteristics. The strain produced the aerial and fragmenting vegetative mycelia consisting of straight chains of 20 or more smooth-surfaced spores. Submerged spores were formed in tryptic soy broth. No soluble pigments were formed. Whole-cell hydrolysates contained glucose and mannose, but not galactose. The 16S rDNA sequence of YC75T was compared with those of the other representative kitasatosporae and streptomycetes. Strain YC75T formed a significant monophyletic clade with Kitasatospora phosalacinea. The levels of DNA relatedness between strain YC75T and representatives of the genus Kitasatospora ranged from 16 to 59% including K. phosalacinea (28 and 40%). It is clear from polyphasic evidence that the isolate should be classified as Kitasatospora cheerisanensis sp. nov., whose type strain is YC75T (= KCTC 2395T). The presence of galactose in whole-cell hydrolysates may not be a stable chemical marker for the genus Kitasatospora.

Thermophilic streptococci were isolated from biofilms on stainless steel samples exposed to pasteurized skimmed milk and from dairy products from a dairy manufacturing plant. The phenotypic characters of these isolates were distinct from those of other thermophilic streptococci of dairy origin (Streptococcus thermophilus and Streptococcus bovis). Genotypic data [restriction endonuclease analysis, ribotyping, random amplified polymorphic DNA (RAPD) profiles, DNA-DNA hybridization and G+C contents] support the classification of these isolates as a new species. The sequence of the 16S rRNA was compared with that of 29 species of streptococci and shown to be significantly different. The sequence of the 16S-23S rRNA intergenic spacer region also differed from published sequences of closely related species. A fluorescent in situ hybridization probe prepared to a specific part of the 16S rRNA gene sequence was able to distinguish the unknown isolates from reference isolates of S. thermophilus and S. bovis. It is proposed that these thermophilic streptococcal isolates from a dairy environment be classified in the genus Streptococcus as a new species, Streptococcus waius (from waiu, the New Zealand Maori word for milk). The type strain is 3/1T(= NZRCC 20100T).

A Gram-positive, aerobic, rod-shaped, peritrichously flagellated, roundendospore-forming bacterium was isolated from a forest soil near Braunschweig, Lower Saxony, Germany, and designated strain HR3-23T(T = type strain). Morphologically, strain HR3-23T shows the characteristics of a member of the genus Bacillus. The spore position is terminal in a swollen sporangium. Comparative analysis of the 16S rDNA sequence shows strain HR3-23T to be most closely related to Caryophanon tenue (95·8% 16S rRNA similarity) and to Bacillus sphaericus (95·4% 16S rRNA similarity).

Phylogenetically, the isolate clusters among species of Bacillus RNA group 2. The DNA G+C content of isolate HR3-23T is 39·3 mol%, the peptidoglycan type is A4α (l-Lys-d-Glu), the major respiratory lipoquinone is menaquinone MK-7 and the predominant fatty acid is of the iso-C15:0 type. Based on the morphological, chemotaxonomic, physiological and phylogenetic properties, a new species, Bacillus silvestris, is proposed; strain HR3-23Tis the type strain (= DSM 12223T).

A Gram-positive, extremely halotolerant bacterium was isolated from the Great Salt Lake, Utah, USA. The strain, designated NNT (= DSM 11805T), was strictly aerobic, rod-shaped, motile by peritrichous flagella and spore-forming. Strain NNT grew at salinities of 0–20% (w/v) NaCI. A distinctive feature of strain NNT was its optimal growth in salt-free medium. The polar lipid pattern of strain NNT consisted of phosphatidyl glycerol, diphosphatidyl glycerol and two phospholipids of unknown structure. The G+C content of its DNA was 38 mol%. The morphological, physiological and, particularly, the 16S rDNA sequence data, showed that strain NNT was associated with ‘Bacillus group 1’. However, the organisms showing the greatest degree of sequence similarity to strain NNT were members of the genus Halobacillus and the species Marinococcus albus, Virgibacillus pantothenticus. Bacillus salexigens and Bacillus dipsosauri. On the basis of chemotaxonomic data, strain NNT was shown to be chemically most similar to B. salexigens and B. dipsosauri, with the greatest degree of similarity being shown to the latter organism. This was consistent with the 16S rDNA sequence data. Members of the genus Halobacillus comprise a chemically distinct group and can easily be distinguished from all other organisms of ‘Bacillus group 1’. On the basis of the 16S rDNA data, chemotaxonomy and the physiology of strain NNT, it is proposed that this organism is a member of a new species, within a new genus, for which the name Gracilibacillus halotolerans is proposed. It is also proposed that B. dipsosauri be transferred to this genus as Gracilibacillus dipsosauri comb. nov. and that B. salexigens be transferred to the genus Salibacillus gen. nov., as Salibacillus salexigens comb. nov. Finally, additional data is provided to support the transfer of Bacillus pantothenticus to the genus Virgibacillus, as Virgibacillus pantothenticus Heyndrickx et al. (1998).

An almost complete gene sequence of 16S rDNA of ‘Nocardia salmonicida’ strain JCM 4826Twas determined following cloning and sequencing of the amplified gene. The sequence was aligned with those available for nocardiae and phylogenetic trees inferred using four tree-making algorithms. The organism and the type strain of Nocardia asteroides consistently formed a monophyletic clade with a distant sequence similarity of 97%. However, previous DNA relatedness experiments showed that strain JCM 4826Tand Nocardia asteroides ATCC 19247Tbelong to different genomic species. The organism was also distinguished from representatives of all validly described species of Nocardia using a combination of phenotypic features. The polyphasic evidence showed that the strain merits recognition as a new species of the genus Nocardia. The name proposed for the new species is Nocardia salmonicida nom. rev.

The genus Actinobispora Jiang et al. is characterized by the formation of longitudinally paired spores on both the vegetative and the aerial mycelium, absence of mycolic acid, cell wall chemotype IV (containing mesodiaminopimelic acid as the diamino acid, galactose and arabinose), PVI and MK-9(H2). Comparative studies of morphology, chemical classification and phylogenetic analysis based on the 16S rRNA sequences among the four type strains of the genus Actinobispora and type strains of related genera were carried out. The results indicated that the genus Actinobispora is different from these other genera and should be placed in the family Pseudonocardiaceae Embley et al. 1988 with the genera Amycolatopsis, Pseudonocardia, Saccharomonospora, Saccharopolyspora and Actinopolyspora. On the bases of these results, three new species under the names of Actinobispora alaniniphila sp. nov., Actinobispora aurantiaca sp. nov. and Actinobispora xinjiangensis sp. nov. are proposed; the type strains are CCTCC AA97001T, CCTCC AA97002T and CCTCC AA97020T.